DJ-1 preserves ischemic postconditioning-induced cardioprotection in STZ-induced type 1 diabetic rats: role of PTEN and DJ-1 subcellular translocation.

BACKGROUND: Ischemic postconditioning (IPostC) has been reported as a promising method for protecting against myocardial ischemia-reperfusion (MI/R) injury. Our previous study found that the infarct-limiting effect of IPostC is abolished in the heart of diabetes whose cardiac expression of DJ-1 (also called PARK7, Parkinsonism associated deglycase) is reduced. However, the role and in particular the underlying mechanism of DJ-1 in the loss of sensitivity to IPostC-induced cardioprotection in diabetic hearts remains unclear. METHODS: Streptozotocin-induced type 1 diabetic rats were subjected to MI/R injury by occluding the left anterior descending artery (LAD) and followed by reperfusion. IPostC was induced by three cycles of 10s of reperfusion and ischemia at the onset of reperfusion. AAV9-CMV-DJ-1, AAV9-CMV-C106S-DJ-1 or AAV9-DJ-1 siRNA were injected via tail vein to either over-express or knock-down DJ-1 three weeks before inducing MI/R. RESULTS: Diabetic rats subjected to MI/R exhibited larger infarct area, more severe oxidative injury concomitant with significantly reduced cardiac DJ-1 expression and increased PTEN expression as compared to non-diabetic rats. AAV9-mediated cardiac DJ-1 overexpression, but not the cardiac overexpression of DJ-1 mutant C106S, restored IPostC-induced cardioprotection and this effect was accompanied by increased cytoplasmic DJ-1 translocation toward nuclear and mitochondrial, reduced PTEN expression, and increased Nrf-2/HO-1 transcription. Our further study showed that AAV9-mediated targeted DJ-1 gene knockdown aggravated MI/R injury in diabetic hearts, and this exacerbation of MI/R injury was partially reversed by IPostC in the presence of PTEN inhibition or Nrf-2 activation. CONCLUSIONS: These findings suggest that DJ-1 preserves the cardioprotective effect of IPostC against MI/R injury in diabetic rats through nuclear and mitochondrial DJ-1 translocation and that inhibition of cardiac PTEN and activation of Nrf-2/HO-1 may represent the major downstream mechanisms whereby DJ-1 preserves the cardioprotective effect of IPostC in diabetes.

Host Gasdermin D restrains systemic endotoxemia by capturing Proteobacteria in the colon of high-fat diet-feeding mice.

Gasdermin D (GSDMD) functions as a key pyroptotic executor through its secreted N-terminal domain (GSDMD-N). However, the functional relevance and mechanistic basis of the precise roles of host colonic GSDMD in high-fat diet (HFD)-induced gut dysbiosis and systemic endotoxemia remain elusive. In this study, we demonstrate that HFD feeding triggers GSDMD-N secretion of both T-lymphocytes and enterocytes in mouse colons. GSDMD deficiency aggravates HFD-induced systemic endotoxemia, gut barrier impairment, and colonic inflammation. More importantly, active GSDMD-N kills the Proteobacteria phylum via directly interacting with Cardiolipin. Mechanistically, we identify that the Glu236 (a known residue for GSDMD protein cleavage) is a bona fide important site for the bacterial recognition of GSDMD. Collectively, our findings explain the mechanism by which colonic GSDMD-N maintains low levels of HFD-induced metabolic endotoxemia. A GSDMD-N mimetic containing an exposed Glu236 site could be an attractive strategy for the treatment of HFD-induced metabolic endotoxemia.

Activation of NRF2/FPN1 pathway attenuates myocardial ischemia-reperfusion injury in diabetic rats by regulating iron homeostasis and ferroptosis.

In patients with ischemic heart disease, myocardial ischemia-reperfusion injury (IRI) can aggravate their condition even worse, and diabetes increases their risk of myocardial IRI. Pathological pathways of common diseases and surgical operations like diabetes, obesity, coronary artery angioplasty, and heart transplantation entail disorders of iron metabolism. Ferroportin1 (FPN1) is the only mammalian protein associated with iron release and thus plays a vital role in iron homeostasis, while nuclear factor E2-related factor 2 (NRF2) controls the transcription of FPN1. Since the NRF2/FPN1 pathway may play a favorable role in the therapy of diabetic myocardial IRI, this work investigated the possible mechanism. In this study, we investigated the effects of ferroptosis in STZ-induced diabetic rats following myocardial IRI in vivo, and its alteration in glucose and hypoxia/reoxygenation-induced cardiomyocytes injury in vitro. Rats and H9c2 cardiomyocytes were randomly divided into 6 groups and treated with sulforaphane and erastin besides the establishment of diabetic myocardial IRI and hyperglycemic hypoxia-reoxygenation models. Cardiac functional and structural damage were detected by Evans blue/TTC double staining, echocardiography, HE staining, and serological indices. CCK-8 assay and ROS production were used to measure cardiomyocyte viability and oxidative stress level. Additionally, the changes in cell supernatant levels of Fe2+, SOD, MDA, and mRNA and protein expression of ferroptosis marker proteins confirmed the beneficial effects of the NRF2/FPN1 pathway on diabetic myocardial IRI related to iron metabolism and ferroptosis. Overall, these findings suggest that iron homeostasis-related ferroptosis plays an important role in aggravating myocardial IRI in diabetic rats, and NRF2/FPN1 pathway-mediated iron homeostasis and ferroptosis might be a promising therapeutic target against myocardial IRI in diabetes.

Mechanism of N-acetylcysteine in alleviating diabetic myocardial ischemia reperfusion injury by regulating PTEN/Akt pathway through promoting DJ-1.

Ischemic heart disease is the main cardiovascular complication of diabetes patients which is mainly caused by oxidative stress. DJ-1 is the key regulator for myocardial protection through inhibiting phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and activating Akt (also known as PKB or protein kinase B). This research is to investigate whether the antioxidant N-acetylcysteine (NAC) could alleviate diabetic myocardial ischemia/reperfusion (I/R) injury by the protective molecule DJ-1. DJ-1 in rat myocardial H9c2 cells and cardiac tissue was respectively knocked down by siRNA and adeno-associated virus (AAV). From the present study, it could be found that compared with high glucose (HG)-normal (N)/DM group, hypoxia/reoxygenation (H/R) or I/R injury can aggravate oxidative stress injury and apoptosis rate of myocardial cells, inhibit the expression of Bcl-2, activate the BAX and cleaved caspase-3 (c-caspase-3) protein and PTEN/Akt pathway. However, in the groups of HG-N, DM, HG-N+I/R and DM+I/R, NAC can significantly reduce oxidative stress injury and apoptosis rate of myocytes, promote the Bcl-2 and DJ-1 molecules, inhibit BAX and c-caspase-3 protein and PTEN/Akt pathway. Compared with HG-N+I/R+NAC and DM+I/R+NAC groups, the oxidative stress injury, apoptosis rate of myocardial cells and heart tissues increased after the knockdown of DJ-1, the expression of Bcl-2 and DJ-1 were inhibited, the BAX and c-caspase-3 expression was increased, and PTEN/Akt pathway was activated. Taken together, the findings suggest that NAC can reduce I/R injury in diabetic myocardium by up-regulating the PTEN/Akt pathway through the level of DJ-1.

Transvaginal Ultrasound-Guided Core Needle Biopsy of Pelvic Masses.

OBJECTIVES: This study assessed the efficacy and safety of transvaginal ultrasound (US)-guided core needle biopsy (CNB) for obtaining adequate pelvic mass samples for histologic analysis and evaluated factors that may affect biopsy success. METHODS: Two hundred cases underwent transvaginal US-guided CNBs for primary inoperable tumors, suspicion of metastases to the ovaries or peritoneum, recurrence, or other solid lesions in the pelvis. Biopsy samples were obtained from the pelvic cavity (67.0%), vaginal cuff or vaginal wall (17.5%), or peritoneal cake (15.5%). The potential influences of the biopsy site (pelvic cavity, vaginal cuff or vaginal wall, or peritoneal cake), vascularization, ascites, tumor size, and tumor type (inoperable, metastases, recurrence, or solid pelvic tumor) on the success of transvaginal US-guided CNB were evaluated by a univariate analysis. RESULTS: Adequate samples were obtained in 192 of 200 biopsies (96.0%), of which 190 yielded successful diagnoses (95.0%). The biopsy site had a significant effect on biopsy adequacy, as there was a significantly lower probability of obtaining satisfactory specimens for histologic verification from the peritoneal cake compared to pelvic tumors and the vaginal cuff or vaginal wall (P < .01). Adequacy was also affected by tumor size (P < .05) but not by vascularization, ascites, or tumor type. No complications occurred during the biopsy procedures. CONCLUSIONS: Transvaginal US-guided CNB is a safe and effective alternative to more invasive methods for evaluating pelvic lesions, such as laparoscopy and laparotomy.

Can quantitative contrast-enhanced ultrasonography predict cervical tumor response to neoadjuvant chemotherapy?

OBJECTIVE: To evaluate the feasibility of quantitative contrast-enhanced ultrasonography (CEUS) for predicting and assessing cervical tumor response to neoadjuvant chemotherapy (NACT). METHODS: Thirty-eight cases with stage IB2 or IIA cervical cancer were studied using CEUS before and after one cycle of NACT. The quantitative CEUS parameters maximum intensity (IMAX), rise time (RT), time to peak (TTP), and mean transit time (MTT) were compared between cervical tumors and myometrium (reference zone) using Sonoliver software. Absolute and relative changes in quantitative CEUS parameters were also compared among complete response, partial response, and non-responsive groups. Correlations between pre-treatment IMAX and changes in quantitative parameters were assessed after one cycle of NACT. RESULTS: There were significant changes in cervical tumor IMAX (P<0.001), RT (P<0.05), and TTP (P<0.05) after one cycle of NACT. According to the Response Evaluation Criteria In Solid Tumors guidelines, the enrollments were divided into complete response, partial response, stable disease and progressive disease groups. There were no significant differences in quantitative CEUS parameters among complete response, partial response, and non-responsive groups (P>0.05). In the stable disease group (n=17), cervical tumor IMAX, RT, and TTP decreased significantly after NACT (P<0.001). The absolute and percentage changes in IMAX were positively correlated with pre-treatment IMAX in all 38 patients (r=0.576, P<0.001 and r=0.429, P<0.001). CONCLUSION: Quantitative CEUS analysis can reveal changes in tumor perfusion following NACT. Tumor perfusion values changes likely precede size changes during the NACT course, and pre-treatment IMAX may be a valuable predictor of cervical tumor perfusion response to NACT with a great decrease in IMAX correlated with better perfusion response.

Contrast-enhanced ultrasonography of cervical carcinoma: perfusion pattern and relationship with tumour angiogenesis.

OBJECTIVE: This study aimed to investigate the use of contrast-enhanced ultrasonography (CEUS) and time-intensity curves to assess angiogenesis in cervical cancer. METHODS: 60 patients who were scheduled to undergo radical surgery for biopsy-proven cervical cancers underwent CEUS. Surgical tissue sections from 32 patients who did not receive neoadjuvant chemotherapy were analyzed with CD34 staining to estimate intratumoral microvessel density (MVD). CEUS images were analyzed for maximum intensity (IMAX), rise time (RT), time to peak (TTP) and mean transit time. RESULTS: Cervical lesions had a higher IMAX and shorter RT and TTP (p < 0.001) than reference regions. There was a linear association between the IMAX of the cervical lesion and the mean intratumoral MVD (r = 0.624, p < 0.001). There were no significant differences in CEUS variables according to histological type, grade and stage. CONCLUSION: Quantitative CEUS variables have potential use for monitoring perfusion changes in tumours after non-surgical therapy for advanced cervical cancer. ADVANCES IN KNOWLEDGE: The article demonstrates the capability and value of quantitative CEUS as a non-invasive strategy for detecting the perfusion and angiogenic status of cervical cancer. Quantitative CEUS variables have potential use for monitoring tumour response to non-surgical therapy.

Reassessment of the predictive role of perivesical fat invasion in invasive bladder cancer prognosis in 151 Chinese patients.

BACKGROUND: Perivesical fat invasion is considered as an important prognostic factor for bladder cancer. However, the predictive role of perivesical fat invasion in invasive bladder cancer prognosis has never been reported in Chinese patients. The aim of the study was to assess the predictive value of perivesical fat invasion for prognoses of T2 and T3 bladder cancer in Chinese patients. METHODS: One hundred and fifty-one patients who underwent radical cystectomy for pT2-3N0M0 invasive bladder cancer from 2001 to 2007 were studied. Cancer-specific survival rate (CSS) and recurrence-free survival rate (RFS) were compared between the pT2 and pT3 patient groups. Other clinicopathological parameters were also retrospectively analyzed by univariate and multivariate analyses to identify the independent predictor for the prognoses of this cohort. RESULTS: Average patient age at surgery was 58 years. Ninety (60.3%) patients had grade I and II disease. During follow-up (median 66 months), 27 patients (17.9%) had tumor recurrence and 18 (11.9%) died of bladder cancer. In the univariate analysis, the CSS and RFS curves between T2 and T3 patients showed no significant difference (P = 0.756 and 0.354, respectively). Multivariate Cox regression showed that histological classification and grade were independent predictors for CSS, while grade was the sole independent predictor for RFS. CONCLUSIONS: For this group of Chinese patients, perivesical fat invasion did not demonstrate a statistically significant difference in prognosis between T2 and T3 patients. Nontransitional cell carcinoma (non-TCC) and high-grade patients had short CSS, and patients with high-grade tumor had higher recurrent risk.

Oncological outcome of surgical treatment in 336 patients with renal cell carcinoma.

BACKGROUND AND OBJECTIVE: The most effective therapy against renal cell carcinoma (RCC) is surgical treatment; however, there have been few large-scale studies that focused on the oncological outcome of this disease in China. The aim of the current study was to report the clinicopathological results and cancer-specific survival (CSS) rate in RCC patients after surgical treatment in our center. METHODS: We retrospectively analyzed the clinicopathological data of 336 RCC patients who underwent radical or partial nephrectomy between 1999 and 2006. Of the 336 patients, 226 were male and 110 were female; the median age was 51 years. Univariate and multivariate analyses were conducted to identify the independent prognostic predictors for this cohort of RCC patients. RESULTS: During follow-up, the overall 5-year CSS rate was 81.4%. The 5-year CSS rates for patients with stage-I, -II, -III, and -IV RCC were 94.7%, 88.9%, 68.8%, and 19.3%, respectively. The patients with T1N0M0 (T1) and T2N0M0 (T2) tumors had similar survival curves. For patients with T1 category tumor, the survival rate did not differ significantly between the radical nephrectomy and nephron-sparing surgery groups. For the 21 patients with metastasis confined to the local lymph nodes, the 5-year survival rate was 31.6% after radical nephrectomy and lymph node dissection. For the 15 patients with vena caval tumor thrombus, the 5-year survival rate was 52.5% after radical nephrectomy and tumor thrombus extirpation. Multivariate Cox regression showed that stage was an independent predictor for CSS (hazard ratio, 3.359; P < 0.001). CONCLUSIONS: For localized RCC, the oncological outcome of this cohort is comparable to that reported in the Western literature. For some patients with locally advanced RCC, aggressive surgical treatment can lead to better long-term survival. However, the prognosis of the patients with metastasis still needs to be improved.

Benign pathological findings in 303 Chinese patients undergoing surgery for presumed localized renal cell carcinoma.

OBJECTIVE: To analyze the incidence of benign lesions in Chinese patients undergoing nephrectomies for renal masses identified as localized renal cell carcinoma (RCC) in preoperative imaging. METHODS: Between 1999 and 2007, 303 patients (112 female, 191 male) with presumed localized RCC underwent nephrectomy (234 radical nephrectomies and 69 partial nephrectomies). Preoperative computed tomography images and pathological findings were reviewed and analyzed. RESULTS: Pathological examinations revealed 31 (10.2%) benign lesions in the 303 patients. Among these 31 benign lesions, 15 (5.0%) were angiomyolipomas (AML) and only four (1.3%) were oncocytomas. Significantly, 20 (17.9%) of the 112 female patients had benign lesions compared with 11 (5.8%; P = 0.001) male patients. Benign renal lesions were found in five (25.0%) of the 20 patients with renal masses smaller than 2 cm, 13 (13.0%) of the 100 patients with renal masses 2-4 cm in size and 13 (7.1%) of the 183 patients with renal masses larger than 4 cm. CONCLUSIONS: Patients in the present study population show a low incidence of benign renal lesions, approximately half of them being AML. Female patients and patients with renal masses smaller than 4 cm are more likely to have benign renal lesions.

A modified technique for neourethral anastomosis in orthotopic neobladder reconstruction.

OBJECTIVES: To introduce a modified technique for urethral anastomosis in orthotopic neobladder reconstruction. METHODS: Between January 2002 and August 2008, about 141 consecutive patients (130 men and 11 women) underwent total cystectomy and orthotopic neobladder reconstruction in which a modified technique was used to anastomosed the caudal-most part of the intestinal neobladder directly to the urethral remnant. The emptying of the neobladder, the early and late complications at the urethral anastomosis was evaluated. RESULTS: Mean patient age at surgery was 58.4 years (range, 33-83) and median follow-up was 42 months (range, 4-83 months). Early urine leakage at the vesicourethral anastomosis developed in 1 patient, and was cured by extending catheter drainage. A total of 138 patients had good emptying of the neobladder, with residual urine volume < 50 mL. Three patients had residual urine volume > 100 mL, and achieved good emptying after intermittent catheterization once a week for 6-12 months. No late complications occurred at the urethral anastomosis site. Daytime continence was good or satisfactory in 97.0% of patients and night-time continence was good or satisfactory in 88.5% of patients. CONCLUSIONS: The clinical outcome of our modified technique for urethral anastomosis in orthotopic neobladder substitution was excellent, but the advantage of this technique needs prospective controlled study.

[Modified intestinal in situ neobladder in functional reconstruction of lower urinary tract after radical cystectomy: report of 15 cases].

BACKGROUND & OBJECTIVE: Quality of life (QOL) in the patients who are performed radical cystectomy is affected by the methods of urinary diversion. The patients with continent or non-continent urinary diversion could not void by themselves and their QOL was poor. However, the intestinal neobladder in situ could provide the patients with good voiding and improve QOL. Unfortunately, some degree of poor empty and incontinence would occur in these patients. The aim of this paper was to report the authors' experience of modified intestinal in situ neobladder in reconstructing the lower urinary tract after radical cystectomy. METHODS: Reconstruction of the lower urinary tract using modified sigmoid neobladder (in 12 patients) and modified ileal neobladder (in 3 patients) was carried out in 15 patients (male 14, female 1; age 33-68 years, mean 53 years) who underwent radical cystectomy for invasive bladder cancer. The patients were followed up for 3-30 months. Nine of them were followed up for more than 16 months. Clinical outcome of these patients including the function of the neobladder, urinary function, sexual status, renal function, serum electrolytes, and QOL was evaluated. RESULTS: All patients voided well by themselves and did not need self-catheterization. Thirteen patients were continent in daytime and night-time. One patient was continent in daytime, but had mild incontinence at night. A female patient had moderate stress incontinence. The capacity of the neobladder was 240-640 ml and the residual volume was 0-250 ml. Renal function was normal in all patients. Serum electrolytes were normal in 14 patients. Metabolic acidosis and unilateral ureteral dilation was encountered in one patient respectively. Nine male patients restored sexual function and 13 patients came back to work. All patients felt satisfied with the function of the neobladder. CONCLUSION: Satisfactory urinary continence and voiding function was achieved with modified intestinal neobladder, which was believed an ideal procedure for lower urinary tract reconstruction after radical cystectomy.